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A microfluidic culture platform for CNS axonal injury, regeneration and transport

Nature Methods volume 2, pages 599–605 (2005)Cite this article

Abstract

Investigation of axonal biology in the central nervous system (CNS) is hindered by a lack of an appropriate in vitro method to probe axons independently from cell bodies. Here we describe a microfluidic culture platform that polarizes the growth of CNS axons into a fluidically isolated environment without the use of targeting neurotrophins. In addition to its compatibility with live cell imaging, the platform can be used to (i) isolate CNS axons without somata or dendrites, facilitating biochemical analyses of pure axonal fractions and (ii) localize physical and chemical treatments to axons or somata. We report the first evidence that presynaptic (Syp) but not postsynaptic (Camk2a) mRNA is localized to developing rat cortical and hippocampal axons. The platform also serves as a straightforward, reproducible method to model CNS axonal injury and regeneration. The results presented here demonstrate several experimental paradigms using the microfluidic platform, which can greatly facilitate future studies in axonal biology.

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Figure 1: The microfluidic-based culture platform directs axonal growth of CNS neurons and fluidically isolates axons.
Figure 2: Axons are isolated without somata or dendrites.
Figure 3: RNA encoding the presynaptic vesicle protein synaptophysin is localized to CNS axons at 6 d in vitro.
Figure 4: Axotomy leads to rapid transcription of immediate early genes and regeneration is enhanced by axonal neurotrophin treatment.
Figure 5: Axons can be cocultured with other cell types such as oligodendrocytes.

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Acknowledgements

We thank W. Poon for help with the radioisotope study. We thank A. Anderson and W. Saadi for reviewing the manuscript. This work was partially funded by the National Institutes of Health (AG-000538, AG-20241, NS-40953 and NS-50895) and the CRPF. A.M.T. thanks the National Institutes of Health for a predoctoral fellowship award (F31NS046208-02).

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Author notes

  1. Anne M Taylor and Mathew Blurton-Jones: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biomedical Engineering, University of California, Irvine, 204 Rockwell Engineering, Irvine, 92697, California, USA

    Anne M Taylor, Seog Woo Rhee & Noo Li Jeon

  2. Institute for Brain Aging and Dementia, University of California, Irvine, 1113 Gillespie NRF, Irvine, 92697, California, USA

    Mathew Blurton-Jones, David H Cribbs & Carl W Cotman

  3. Department of Neurology, University of California, Irvine, 1207 Gillespie NRF, Irvine, 92697, California, USA

    David H Cribbs & Carl W Cotman

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  1. Anne M Taylor
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Correspondence to Noo Li Jeon.

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The authors have applied for a patent which covers the device described in the article.

Supplementary information

Supplementary Fig. 1

The microfluidic platform facilitates live imaging of axonal transport. (PDF 55 kb)

Supplementary Methods (PDF 53 kb)

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Taylor, A., Blurton-Jones, M., Rhee, S. et al. A microfluidic culture platform for CNS axonal injury, regeneration and transport. Nat Methods 2, 599–605 (2005). https://doi.org/10.1038/nmeth777

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